Bioinformatics approach to understand the mode of microbial pathogenesis of Chlamydia trachomatis and their implications in gynecologic malignancy.

Chlamydia trachomatis has a say on the target gene i.e., modulating the expression of target gene in the host so that it is given protection from the immune cells and so its survival and replication are not arrested by the host. The current study reports a wide range of C. trachomatis proteins that target the cellular as well as sub-cellular components of the host in gynecologic malignancy. Various bioinformatics tools was used to conduct an in-depth analysis on nuclear and eukaryotic sub cellular localization signal to find the sequences of the predicted proteins of C. trachomatis strain G. A total of 411 proteins was identified with 79.54% maximum expected accuracy and 51.02% least expected accuracy. There were uneven prediction of proteins along with redundancies between BaCeILo and HSLpred in the determination of sub-cellular localization of the CT proteins. The highest molecular weight proteins (>80 kDa) were observed to be the targeted proteins to nucleus of host cell. There was no constant patterns observed in the values of isoelectric point (pI) in case of mitochondrial targeting. The expression of eight proteins were significant with different fold changes. The in-silico study provided much detailed insights for further research in gynecological cancer. However, further experiments should be conducted to validate the specificity and confirmatory roles played by these predicted proteins in carcinogenesis.

Oncogene mutational analysis in imatinib naive population of gastrointestinal stromal tumor patients.

There are very scanty reports on gastro-intestinal stromal tumors (GISTs), a very common tumor of mesenchymal cells in GIT track primary resistance to imatinib. This comprehensive study identifies the prevalence, clinical presentation and GIST genotype association in the imatinib naïve population. Prospectively a record of anthropometric, baseline demographic data and clinical details for the patients diagnosed with GIST were scrutinized. Pathological information included the presence or absence of necrosis, tumor size, mitotic counts, immune-histochemical staining for CD 34, CD 117 and DOG1 was performed using biopsy sample. Selected exon genes of KIT, PDGFRA and BRAF were amplified and subjected to mutation analysis by direct sequencing. Appropriate statistical analyses were performed. The male/female ratio was 1.8:1 among 54 patients with GIST. The mean GIST size was comparatively bigger in females (2.49±0.855) than males (2.26±1.13). The stomach was the most common site for GIST followed by the Small bowel and rectum. The majority of the tumours were spindle cell. This study reports 12 different types of mutation among 39 KIT, 8 PDGFRA and 7 BRAF mutations. In KIT, the most prevalent was exon 11 mutation with the KITdelinc557/558 (14/30) being the major exon 11 type mutation. In PDGRFA, five exons 18 with p.D842V substitution and three exons 12 deletion mutation was reported. Seven patients had strong or diffuse BRAF staining having V600E type mutation as major BRAF type mutation. Drug-resistant GIST due to acquired mutations remains a serious issue, therefore genetic information of such mutational related to drug-resistant may provide the imperative clue for diagnosis and clinical treatment. These mutations are pivotal for prognosis and associated with imatinib as not all of them but only a few are reported resistant to the imatinib.

Modulation of host immune status by cryptococcus co-infection during HIV-1 pathogenesis and its impact on CD+4 cell and cytokines environment.

BACKGROUND: Cryptococcus infection is the second most common opportunistic infection in HIV patients with an increased rate of morbidity and mortality. Altered immune system during HIV- Cryptococcus co-infection is yet to be explored by laboratory. This study evaluates pro- and the anti-inflammatory cytokines in HIV patients with Cryptococcus co-infection and correlate them with CD4+T cell counts as well as viral loads before the initiation of drug therapy. This information would enable to understand host immune modulation and cellular environment during co-infection and understand its impact on HIV pathogenesis. METHODOLOGY: The study comprised four categories of patients with cryptococcosis, HIV, HIV-cryptococcosis co-infected and asymptomatic Healthy volunteers. All the patients and healthy individuals were subjected to CD4+T cells count by FACS using monoclonal antibody cocktail CD4+T cell count (counts per mm3) which was counted using multiSET software on FACS caliber. The viral loads were counted in terms of viral RNA copy numbers which was estimated by real-time PCR using by Artus HIV-1 RG. The sensitivity of kit was >70 IU/ml. ELISA was performed for IL-12 p70, IL-12, IL-4, IL-10, IL-6, TNF-α and IFN-Y using commercially kits (BD Biosciences, USA). Significant variations were assayed by Student's t-test and P values ≤ 0.05 were considered statistically significant. RESULTS: Reduction in CD+4 cell counts was highly significant in HIV patients with or without cryptococcosis. CD4+T cell counts were inversely proportional to viral load. TNF-α levels were raised in cryptococcosis patients significantly higher than healthy individuals. TNF-α was more or less not dependent on viral load but it was more related to the cryptococcosis IL-12 levels were increased in patients with infection and was highest in the HIV infected group. Level of IL-4 was similar in healthy and patients with cryptococcosis but it was elevated in HIV-Crypto co-infected patients. HIV infected patients showed a significant increase in IL-4 level and it was elevated higher in co-infected patients. IL-10 and IL-6 were significantly higher in HIV patients. The fungal infection did not influence the levels of IL-10 in HIV group but IL-6 was low in fungal infected patients. CONCLUSION: There are very limited studies related to the immune modulation status of HIV co-infected with Cryptococcus before the initiation of any drug therapy. Such information might through in-depth light to understand the initial state of the immune environment which certainly would play a pivotal role in the outcome of the immune modulation.